UNIVERSITY  OF  CALIFORNIA 
AT  LOS  ANGELES 


GIFT  OF  CAPT.  AND  MRS. 
PAUL  MCBRIDE  PERIGORD 


ELEMENTARY  TECHNIQUE 


IN 


HISTOLOGY  AND  BACTERIOLOGY 


BY 

ERNEST  B.    HOAG,   A.  B.,  B.  S. 

Instructor  in  Zoology  and  Physiology, 

THROOP  POLYTECHNIC  INSTITUTE, 

Pasadena,  Cal. 


H.    KAHN,    PHAR.   M. 

Assistant  Demonstrator  in   Bacteriology, 

NORTHWESTERN  UNIVERSITY  MEDICAL  SCHOOL, 

Chicago. 


CHICAGO: 

THE  W.  T.   KEENER  CO. 
1896. 

136516 


Entered  according  to  Act  of  Congress,  in  the  year  1895,  by 

Ernest  B.  Hoag  and  H.  Kahn, 
in  the  Office  of  the  Librarian  of  Congress,  at  Washington. 


5.5-.  5- 


PREFACE. 


This  guide  in  elementary  technique  is  intended  for 
the  use  of  medical  and  other  students  beginning 
work  in  either  Histology  or  Bacteriology. 


ERRATA. 

Page  12.  19,  :23,  for  "creasote"  read  "creosote." 

Page  15,  line  3,  for  "tissues"  read  "tissue." 

Page  40,  for  "Shafer"  read  -'Sehafer." 

Page  16,  17,  20  for  "flxitive"  read  "fixative." 

Page  71,  eighth  line  from  bottom,  for  "or  covered"  read  "are  covered." 

For  "liquifying"  read  ''liquefying." 


are  usuany  taxen  ror  granted,  ana  wmcn  tneretore,  often" 
greatly  perplex  the  student. 

It  is  not  expected  that  the  book  will  furnish  the  ex- 
perienced student  with  much  information;  it  is  written 
to  help  the  beginner  and  while  it  is  thought  that  it  will 
be  sufficiently  complete  for  the  requirements  of  all  such, 
we  especially  hope  that  it  will  serve  as  an  introduction 
to  more  advanced  works,  and  that  the  result  will  be, 
a  clear  understanding  of  the  elementary  methods,  and 
therefore  a  much  more  intelligent  use  of  the  advanced 
books  on  Histology  and  Bacteriology. 

It  will  be  noted,  that  no  attempt  has  been  made  to 
introduce  descriptive  Histology  and  scarcely  any  de- 
scriptive Bacteriology,  but  that  the  book  is  confined  to 
Technique. 


S5".  5" 


PREFACE. 


This  guide  in  elementary  technique  is  intended  for 
the  use  of  medical  and  other  students  beginning 
work  in  either  Histology  or  Bacteriology. 

In  its  main  features  it  is  a  new  arrangement 
of  what  is  found  in  other  books  on  these  subjects,  and 
in  many  instances  direct  quotations  are  made. 

The  absence  of  any  concise,  and  yet  sufficiently  full, 
account  of  methods  which  will  enable  the  beginner  to 
form  from  the  first,  clear  ideas  of  the  various  steps  is 
the  only  apology  offered  for  adding  another  book  to  the 
many  excellent  ones  now  to  be  found. 

It  is  hoped  that  this  little  book  will  answer  that  com  - 
mon  question  of  the  beginner,  "What  shall  I  do  next:"' 
We  have  tried  to  make  plain  those  simple  points  which 
are  usually  taken  for  granted,  and  which  therefore,  often 
greatly  perplex  the  student. 

It  is  not  expected  that  the  book  will  furnish  the  ex- 
perienced student  with  much  information;  it  is  written 
to  help  the  beginner  and  while  it  is  thought  that  it  will 
be  sufficiently  complete  for  the  requirements  of  all  such, 
we  especially  hope  that  it  will  serve  as  an  introduction 
to  more  advanced  works,  and  that  the  result  will  be, 
a  clear  understanding  of  the  elementary  methods,  and 
therefore  a  much  more  intelligent  use  of  the  advanced 
books  on  Histology  and  Bacteriology. 

It  will  be  noted,  that  no  attempt  has  been  made  to 
introduce  descriptive  Histology  and  scarcely  any  de- 
scriptive Bacteriology,  but  that  the  book  is  confined  to 
Technique. 


PREFACE. 

We  are  particularly  indebted  for  valuable  assistance 
received  from  Dr.  Stanley  P.  Black,  Pathologist  at 
Mercy  Hospital.  Dr.  F.  X.  Walls,  and  Dr.  E.G.  Conklin 
of  Northwestern  University,  Dr.  A.  C.  Eychleshymer 
of  Chicago  University,  and  Dr  Adolph  Gehrmann  of 
College  of  Physicians  and  Surgeons,  Chicago,  have  also 
rendered  us  much  assistance. 

Among  the  books  consulted,  we  have  drawn  more 
particularly  for  Part  I  from  Lee's  Vade  Mecum,  Stir- 
ling's Histology,  Schafer's  Histology,  Von  Kahlden's 
Pathological  Histology,  and  Lehrbuch  der  Histologie 
und  Mikroskopischen  Techink.  (Bohm  und  Davidoff), 
and  for  Part  II  from  Migula,  Sternberg,  Schenk,  Novy, 
and  Praenkel.  Suggestions  and  criticisms  from  those 
interested  in  the  material  here  presented,  will  be  greatly 
appreciated. 

September,  1895. 


PART  I. 

HISTOLOGY. 


HISTOLOGY  AND  BACTERIOLOGY.  5 

2nd.  Hardening  the  elements  of  a  tissue  so  that  their 
structure  will  remain  as  nearly  normal  in  appearance  as 
possible,  after  the  various  reagents  for  their  preparation 
have  been  used. 

The  fixing  of  a  tissue  is  of  the  greatest  importance, 
and  final  success  or  failure  with  the  sections  depends 
very  largely  upon  this  step. 

One  must  know  what  fixing  agent  to  use  for  a  par- 
ticular tissue;  how  long  to  allow  it  to  act;  with  what  to 
wash  out  the  fluid,  and  what  strength  of  alcohol  to  use 
after  washing. 

Secure  perfectly  fresh  tissue  if  possible.  Use 
small  pieces,  and  15  to  20  times  their  volume  of  the 
fixing  fluid. 

There  are  many  good  fixing  agents  and  the  choice  of 
one  depends  upon  the  kind  of  tissue  and  the  result  de- 
sired. For  most  purposes  the  following  will  be  found 
satisfactory  fixing  fluids,  and  among  them,  corrosive 
sublimate,  absolute  alchohol,  Flemming's  fluid  and  Per- 
enyi's  fluid  are  particularly  recommended. 

FIXING  AGENTS  IN  GENERAL  USE. 

1.     PERENYI'S  FLUID.— Formula  on  page  32. 

Objects  are  left  in  the  fluid  from  3  to  5  hours  for  small 
embryos  and  4  to  12  hours  for  the  tissues  of  vertebrates, 
and  then  transferred  directly  to  70-75  per  cent,  alcohol 
for  at  least  24  hours.  They  may  then  be  placed  in  80 
per  cent  alcohol  and  left  in  this  until  wanted,  or  they 
may  be  dehydrated  at  once  by  carrying  them  through 
the  alcohols,  including  absolute.  This  is  a  very  valua- 
ble fluid  for  both  embryonic  and  adult  tissues.  No  seri- 
ous results  follow  when  a  tissue  is  left  in  the  fluid  for  a 
number  of  hours.  Borax  carmine  may  be  added  to  70 
per  cent  alcohol  so  that  the  hardening  and  staining  is 
done  at  the  same  time. 


HISTOLOGY  AND  BACTERIOLOGY.  5 

2nd.  Hardening  the  elements  of  a  tissue  so  that  their 
structure  will  remain  as  nearly  normal  in  appearance  as 
possible,  after  the  various  reagents  for  their  preparation 
have  been  used. 

The  fixing  of  a  tissue  is  of  the  greatest  importance, 
and  final  success  or  failure  with  the  sections  depends 
very  largely  upon  this  step. 

One  must  know  what  fixing  agent  to  use  for  a  par- 
ticular tissue;  how  long  to  allow  it  to  act;  with  what  to 
wash  out  the  fluid,  and  what  strength  of  alcohol  to  use 
after  washing. 

Secure  perfectly  fresh  tissue  if  possible.  Use 
small  pieces,  and  15  to  20  times  their  volume  of  the 
fixing  fluid. 

There  are  many  good  fixing  agents  and  the  choice  of 
one  depends  upon  the  kind  of  tissue  and  the  result  de- 
sired. For  most  purposes  the  following  will  be  found 
satisfactory  fixing  fluids,  and  among  them,  corrosive 
sublimate,  absolute  alchohol,  Flemming's  fluid  and  Per- 
enyi's  fluid  are  particularly  recommended. 

FIXING  AGENTS  IN  GENERAL  USE. 

1.     PERENYI'S  FLUID. — Formula  on  page  32. 

Objects  are  left  in  the  fluid  from  3  to  5  hours  for  small 
embryos  and  4  to  12  hours  for  the  tissues  of  vertebrates, 
and  then  transferred  directly  to  70-75  per  cent,  alcohol 
for  at  least  24  hours.  They  may  then  be  placed  in  80 
per  cent  alcohol  and  left  in  this  until  wanted,  or  they 
may  be  dehydrated  at  once  by  carrying  them  through 
the  alcohols,  including  absolute.  This  is  a  very  valua- 
ble fluid  for  both  embryonic  and  adult  tissues.  No  seri- 
ous results  follow  when  a  tissue  is  left  in  the  fluid  for  a 
number  of  hours.  Borax  carmine  may  be  added  to  70 
per  cent  alcohol  so  that  the  hardening  and  staining  is 
done  at  the  same  time. 


